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Rhombomere-specific patterns of apoptosis in the tree shrew Tupaia belangeri

Abstract

Whether rhombomere-specific patterns of apoptosis exist in the developing hindbrain of vertebrates is under debate. We have investigated the sequence of apoptotic events in three-dimensionally reconstructed hindbrains of Tupaia belangeri (8- to 19-somite embryos). Apoptotic cells were identified by structural criteria and by applying an in situ tailing technique to visualize DNA fragmentation. Seven rhombomeres originated from three pro-rhombomeres. Among pre-migratory neural crest cells in the dorsal thirds of the neural folds, the earliest apoptotic concentrations appeared in the developing third rhombomere (r3). Dorsal apoptotic maxima then persisted in r3, extended from r3 to r2, and also arose in r5. Transverse apoptotic bands increased the total amount of apoptotic cells in odd-numbered rhombomeres first in r3 and, with a delay, also in r5. This sequence of apoptotic events was paralleled by an approximate rostrocaudal sequence of neural crest cell delamination from the even-numbered rhombomeres. Thus, large-scale apoptosis in r3 and r5 helped to establish crest-free zones that segregated streams of migrating neural crest cells adjacent to r2, r4, and r6. The sequence of apoptotic events observed in the dorsal thirds of rhombomeres matches that reported for the chick embryo. Other shared features are apoptotic peaks in the position of a circumscribed ventricular protrusion of fusing parts of the neural folds in r1 and r2, and Y-shaped apoptotic patterns composed of apoptotic maxima in the dorsal and lateral thirds of r1, r2, and r3. These rhombomere-specific patterns of apoptosis may therefore represent a conserved character, at least in amniotes.

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References

  1. Barrow JR, Stadler HS, Capecchi MR (2000) Roles of Hoxa1 and Hoxa2 in patterning the early hindbrain of the mouse. Development 127:933–944

  2. Birgbauer E, Fraser SE (1994) Violation of cell lineage restriction compartments in the chick hindbrain. Development 120:1347–1356

  3. Birgbauer E, Sechrist J, Bronner-Fraser M, Fraser S (1995) Rhombomeric origin and rostrocaudal reassortment of neural crest cells revealed by intravital microscopy. Development 121:935–945

  4. Boissonnat JD (1988) Shape reconstruction from planar cross sections. Comput Vision Graph Image Proc 44:1–29

  5. Brunnett G, Vanco M, Haller C, Washausen S, Kuhn H-J, Knabe W (2003) Visualization of cross sectional data for morphogenetic studies. In: Dittrich K, König W, Oberweis A, Rannenberg K, Wahlster W (eds) Proceedings GI Workshop “Visualisierung in der Bioinformatik”. Lecture notes in informatics, vol 34. Köllen, Bonn, pp 354–359

  6. Cooke JE, Moens CB (2002) Boundary formation in the hindbrain: Eph only it were simple... . Trends Neurosci 25:260–267

  7. Duboule D, Dolle P (1989) The structural and functional organization of the murine HOX gene family resembles that of Drosophila homeotic genes. EMBO J 8:1497–1505

  8. Eickholt BJ, Graham A, Lumsden A, Wizenmann A (2001) Rhombomere interactions control the segmental differentiation of hindbrain neurons. Mol Cell Neurosci 18:141–148

  9. Ellies DL, Church V, Francis-West P, Lumsden A (2000) The WNT antagonist cSFRP2 modulates programmed cell death in the developing hindbrain. Development 127:5285–5295

  10. Farlie PG, Kerr R, Thomas P, Symes T, Minichiello J, Hearn CJ, Newgreen D (1999) A paraxial exclusion zone creates patterned cranial neural crest cell outgrowth adjacent to rhombomeres 3 and 5. Dev Biol 213:70–84

  11. Fraser S, Keynes R, Lumsden A (1990) Segmentation in the chick embryo hindbrain is defined by cell lineage restrictions. Nature 344:431–435

  12. Goping G, Wood KA, Sei Y, Pollard HB (1999) Detection of fragmented DNA in apoptotic cells embedded in LR White: a combined histochemical (LM) and ultrastructural (EM) study. J Histochem Cytochem 47:561–568

  13. Graham A, Papalopulu N, Krumlauf R (1989) The murine and Drosophila homeobox gene complexes have common features of organization and expression. Cell 57:367–378

  14. Graham A, Heyman I, Lumsden A (1993) Even-numbered rhombomeres control the apoptotic elimination of neural crest cells from odd-numbered rhombomeres in the chick hindbrain. Development 119:233–245

  15. Graham A, Francis-West P, Brickell P, Lumsden A (1994) The signalling molecule BMP4 mediates apoptosis in the rhombencephalic neural crest. Nature 372:684–686

  16. Graham A, Koentges G, Lumsden A (1996) Neural crest apoptosis and the establishment of craniofacial pattern: an honorable death. Mol Cell Neurosci 8:76–83

  17. Häcker G (2000) The morphology of apoptosis. Cell Tissue Res 301:5–17

  18. Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92

  19. Heyman I, Faissner A, Lumsden A (1995) Cell and matrix specialisations of rhombomere boundaries. Dev Dyn 204:301–315

  20. Heyman I, Kent A, Lumsden A (1993) Cellular morphology and extracellular space at rhombomere boundaries in the chick embryo hindbrain. Dev Dyn 198:241–253

  21. Jeffs P, Jaques K, Osmond M (1992) Cell death in cranial neural crest development. Anat Embryol 185:583–588

  22. Knabe W, Kuhn H-J (1998) Pattern of cell death during optic cup formation in the tree shrew Tupaia belangeri. J Comp Neurol 401:352–366

  23. Knabe W, Kuhn H-J (1999) The earliest invasion of macrophages into the developing brain and eye of the tree shrew Tupaia belangeri. Anat Embryol 200:393–402

  24. Knabe W, Süss M, Kuhn H-J (2000) The patterns of cell death and of macrophages in the developing forebrain of the tree shrew Tupaia belangeri. Anat Embryol 201:157–168

  25. Knabe W, Washausen S, Brunnett G, Kuhn H-J (2002) Use of “reference series” to realign histological serial sections for three-dimensional reconstructions of the positions of cellular events in the developing brain. J Neurosci Methods 121:169–180

  26. Köntges G, Lumsden A (1996) Rhombencephalic neural crest segmentation is preserved throughout craniofacial ontogeny. Development 122:3229–3242

  27. Kuhn H-J, Schwaier A (1973) Implantation, early placentation, and the chronology of embryogenesis in Tupaia belangeri. Anat Embryol 142:315–340

  28. Kuratani S (1997) Spatial distribution of postotic crest cells defines the head/trunk interface of the vertebrate body: embryological interpretation of peripheral nerve morphology and evolution of the vertebrate head. Anat Embryol 195:1–13

  29. Lawson A, England MA (1998) Neural fold fusion in the cranial region of the chick embryo. Dev Dyn 212:473–481

  30. Lawson A, Schoenwolf GC, England MA, Addai FK, Ahima RS (1999) Programmed cell death and the morphogenesis of the hindbrain roof plate in the chick embryo. Anat Embryol 200:509–519

  31. Lumsden A (1990) The cellular basis of segmentation in the developing hindbrain. Trends Neurosci 13:329–335

  32. Lumsden A (1999) Closing in on rhombomere boundaries. Nat Cell Biol 1:E83–E85

  33. Lumsden A, Keynes R (1989) Segmental patterns of neuronal development in the chick hindbrain. Nature 337:424–428

  34. Lumsden A, Sprawson N, Graham A (1991) Segmental origin and migration of neural crest cells in the hindbrain region of the chick embryo. Development 113:1281–1291

  35. Moens CB, Prince VE (2002) Constructing the hindbrain: insights from the zebrafish. Dev Dyn 224:1–17

  36. Osumi-Yamashita N, Ninomiya Y, Doi H, Eto K (1996) Rhombomere formation and hind-brain crest cell migration from prorhombomeric origins in mouse embryos. Dev Growth Differ 38:107–118

  37. Peterson PE, Blankenship TN, Wilson DB, Hendrickx AG (1996) Analysis of hindbrain neural crest migration in the long-tailed monkey (Macaca fascicularis). Anat Embryol 194:235–246

  38. Romeis B (1989) Heidenhain’s hematoxylin. In: Böck P (ed) Mikroskopische Technik, 17th edn. Urban-Schwarzenberg, Baltimore, pp 10-100

  39. Ruberte E, Wood HB, Morriss-Kay GM (1997) Prorhombomeric subdivision of the mammalian embryonic hindbrain: is it functionally meaningful? Int J Dev Biol 41:213–222

  40. Sanders EJ, Wride MA (1995) Programmed cell death in development. Int Rev Cytol 163:105–173

  41. Sechrist J, Nieto MA, Zamanian RT, Bronner-Fraser M (1995) Regulative response of the cranial neural tube after neural fold ablation: spatiotemporal nature of neural crest regeneration and up-regulation of Slug. Development 121:4103–4115

  42. Serbedzija GN, Bronner-Fraser M, Fraser SE (1992) Vital dye analysis of cranial neural crest cell migration in the mouse embryo. Development 116:297–307

  43. Serbedzija GN, Dickinson M, McMahon AP (1996) Cell death in the CNS of the Wnt-1 mutant mouse. J Neurobiol 31:275–282

  44. Smith A, Graham A (2001) Restricting BMP-4 mediated apoptosis in hindbrain neural crest. Dev Dyn 220:276–283

  45. Smith A, Robinson V, Patel K, Wilkinson DG (1997) The EphA4 and EphB1 receptor tyrosine kinases and ephrin-B2 ligand regulate targeted migration of branchial neural crest cells. Curr Biol 7:561–570

  46. Süss M, Washausen S, Kuhn H-J, Knabe W (2002) High resolution scanning and three-dimensional reconstruction of cellular events in large objects during brain development. J Neurosci Methods 113:147–158

  47. Tan SS, Morriss-Kay G (1985) The development and distribution of the cranial neural crest in the rat embryo. Cell Tissue Res 240:403–416

  48. Trainor PA, Sobieszczuk D, Wilkinson D, Krumlauf R (2002) Signalling between the hindbrain and paraxial tissues dictates neural crest migration pathways. Development 129:433–442

  49. Weil M, Jacobson MD, Raff MC (1997) Is programmed cell death required for neural tube closure? Curr Biol 7:281–284

  50. Wible JR, Zeller U (1994) Cranial circulation of the pen-tailed tree shrew Ptilocercus lowii and relationships of Scandentia. J Mamm Evol 2:209–230

  51. Wilkinson DG, Bhatt S, Cook M, Boncinelli E, Krumlauf R (1989) Segmental expression of Hox-2 homeobox-containing genes in the developing mouse hindbrain. Nature 341:405–409

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Acknowledgements

We thank Dr. Marek Vanco, Technische Universität Chemnitz, for help with the three-dimensional reconstructions, and Irmgard Weiß for technical assistance.

Author information

Correspondence to Wolfgang Knabe.

Additional information

This work was supported by the Deutsche Forschungsgemeinschaft (KN 525/1–1, KN 525/1–2, BR 1185/4–1, and former Sonderforschungsbereich 89: Cardiology)

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Knabe, W., Washausen, S., Brunnett, G. et al. Rhombomere-specific patterns of apoptosis in the tree shrew Tupaia belangeri . Cell Tissue Res 316, 1–13 (2004). https://doi.org/10.1007/s00441-004-0855-0

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Keywords

  • Hindbrain
  • Development
  • Cell death
  • Neural crest cells
  • Tree shrew, Tupaia belangeri
  • Scandentia